非小细胞肺癌中免疫检查点及免疫治疗中细胞亚群的异质性  

Study on the Heterogeneity of Cell Subpopulations in Immunotherapy and Immune Checkpoints in Non-small Cell Lung Cancer

作　　者：杨燕 陈丹[2] 朱红革[1] Yang Yan;Chen Dan;Zhu Hongge(Department of Pulmonary Medicine,Cancer Hospital Affiliated to Xinjiang Medical University,Urumqi 830011;School of Public Health,Xinjiang Medical University,Urumqi 830054)

机构地区：[1]新疆医科大学附属肿瘤医院肺内科,乌鲁木齐830011 [2]新疆医科大学公共卫生学院,乌鲁木齐830054

出　　处：《国际老年医学杂志》2024年第6期653-661,共9页International Journal of Geriatrics

基　　金：新疆维吾尔自治区自然科学基金(2022D01C796,2022D01C523)。

摘　　要：目的 鉴定非小细胞肺癌(NSCLC)中分子失调机制、免疫检查点异常以及反应于免疫治疗的免疫细胞。方法 收集GSE81089,GSE120622和TCGA数据库中NSCLC和对照的基因表达数据并进行差异表达和富集分析。鉴定差异表达的免疫检查点对总体生存的影响。收集2022年6月—2023年5月于新疆医科大学附属肿瘤医院接受新辅助治疗的NSCLC患者的肿瘤和癌旁对照组织以及血液样本,利用qRT-PCR检测肿瘤和癌旁对照组织中免疫检查点的表达。另外,在GSE207422单细胞数据集中鉴定新辅助治疗后主要病理缓解(MPR)和非主要病理缓解(NMPR)之间免疫细胞的差异,进一步通过流式细胞术检测MPR和NMPR之间免疫细胞的丰度差异。结果 在GSE81089,GSE120622和TCGA中鉴定了2 505个差异表达基因的交集,富集分析发现交集基因主要参与神经活性配体-受体相互作用、PI3K-Akt信号通路和细胞因子-细胞因子受体相互作用等信号通路;此外,有6个免疫检查点在NSCLC中差异表达,CD40LG、CD160、VTCN1、TDO2显著影响患者的总体生存。与癌旁对照组比较,qRT-PCR检测证实了CD40LG和CD160在NSCLC中显著低表达(P<0.05),而VTCN1和TDO2显著高表达(P<0.05)。通过整合GSE207422单细胞数据,将32个细胞群分为13种细胞亚型,其中CD4、CD8、B细胞、NK和M2巨噬细胞在MPR中明显富集,流式细胞术检测证实了这些免疫细胞在MPR中的丰度明显高于NMPR。结论 免疫检查点与NSCLC患者总体生存显著相关,并确认了MPR中富集的免疫细胞。这些发现为NSCLC的免疫治疗提供了新的生物学见解和潜在的治疗靶点。Objective To identify molecular dysregulation mechanisms,immune checkpoint abnormalities,and immune cells responsive to immunotherapy in non-small cell lung cancer(NSCLC).Methods Gene expression data for NSCLC and controls from the GSE81089,GSE120622,and TCGA databases were collected and subjected to differential expression and enrichment analysis.The impact of differentially expressed immune checkpoints on overall survival was identified.Tumor tissues,adjacent control tissues,and blood samples were collected from NSCLC patients who received neoadjuvant therapy at Cancer Hospital Affiliated to Xinjiang Medical University from June 2022 to May 2023,the expression of immune checkpoints in tumor and adjacent control tissues was detected using qRT-PCR.Additionally,in the GSE207422 single-cell dataset,differences in immune cells between major pathological remission(MPR)and non-major pathological remission(NMPR)after neoadjuvant therapy were identified.The abundance differences of immune cells between MPR and NMPR were further detected by flow cytometry.Results In the GSE81089,GSE120622,and TCGA datasets,an intersection of 2505 differentially expressed genes was identified.Enrichment analysis revealed that these intersecting genes are primarily involved in neuroactive ligand-receptor interaction,the PI3K-Akt signaling pathway,and cytokine-cytokine receptor interaction,among other signaling pathways.Additionally,six immune checkpoints were differentially expressed in NSCLC,with CD40LG,CD160,VTCN1,and TDO2 significantly impacting overall patient survival.Compared to the control group,qRT-PCR confirmed that CD40LG and CD160 were significantly downregulated in NSCLC(P<0.05),while VTCN1 and TDO2 were significantly upregulated(P<0.05).By integrating the GSE207422 single-cell dataset,32 cell clusters were classified into 13 cell subtypes.Comparisons revealed a significant enrichment of CD4,CD8,B cells,NK cells,and M2 macrophages in MPR.Flow cytometry confirmed that the abundance of these immune cells was higher in MPR than in

关 键 词：非小细胞肺癌 免疫检查点 主要病理缓解 免疫细胞 

分 类 号：R734.2[医药卫生—肿瘤]